Immunoprecipitation was performed on 3 mg of protein with RON c-terminal (E9, Santa Cruz) and RON N-terminal (RON8, Imclone) then beads were submitted and sample preparation performed as previously described 41 (link). Trypsin-digested peptides were analyzed by high pressure liquid chromatography (HPLC) coupled with tandem mass spectroscopy (LC-MS/MS) using nano-spray ionization 42 (link). The nanospray ionization experiments were performed using a TripleTof 5600 hybrid mass spectrometer (ABSCIEX) interfaced with nano-scale reversed-phase HPLC (Tempo). The collected data were analyzed using MASCOT® (Matrix Sciences) and Protein Pilot 4.0 (ABSCIEX) for peptide identifications. RON isoform protein sequences are not normally included in the Protein Pilot analysis software and were supplemented in order to determine their detection.
Tripletof 5600 hybrid mass spectrometer
Manufactured by AB Sciex
The TripleTof 5600 hybrid mass spectrometer is a laboratory instrument designed for high-resolution, accurate mass measurements of a wide range of analytes. It combines the features of a triple quadrupole and a time-of-flight mass analyzer, providing a versatile platform for various analytical applications.
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Lab products found in correlation
3 protocols using tripletof 5600 hybrid mass spectrometer
1
Proteomic Analysis of RON Isoforms
2
Proteomic Analysis of RON Isoforms
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Immunoprecipitation was performed on 3 mg of protein with RON c-terminal (E9, Santa Cruz) and RON N-terminal (RON8, Imclone) then beads were submitted and sample preparation performed as previously described 41 (link). Trypsin-digested peptides were analyzed by high pressure liquid chromatography (HPLC) coupled with tandem mass spectroscopy (LC-MS/MS) using nano-spray ionization 42 (link). The nanospray ionization experiments were performed using a TripleTof 5600 hybrid mass spectrometer (ABSCIEX) interfaced with nano-scale reversed-phase HPLC (Tempo). The collected data were analyzed using MASCOT® (Matrix Sciences) and Protein Pilot 4.0 (ABSCIEX) for peptide identifications. RON isoform protein sequences are not normally included in the Protein Pilot analysis software and were supplemented in order to determine their detection.
3
Proteomic Analysis of Peritoneal Cells
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CD-1 mice were injected i.p. with 400 µg of POPC liposomes and total peritoneal cells were collected 1 h later by peritoneal lavage, centrifuged, and cell pellets were processed for mass spectrometry analysis by the Biomolecular and Proteomics Mass Spectrometry Facility (University of California, San Diego). Control animals received an equal volume of PBS. Briefly, samples were digested with trypsin and analyzed by HPLC coupled with tandem mass spectrometry (LC-MS/MS) using nano-spray ionization (TripleTOF 5600 hybrid mass spectrometer (AB SCIEX)). MS/MS spectra acquisition, peptide/protein identification, quantification of the data, and statistical calculations were performed using the peptide-feature-based PEAKS Studio X (Bioinformatics Solutions Inc., Canada) [12 (link)].
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